Gene expression
BMDM, THP-1 cells, or human monocytes were cultured with 200 ng/ml LPS for 4 hours and then treated with 10 or 100 μM LMT503 for 18 hours at 37°C. To investigate gene expression in colon macrophages, mononuclear cells were isolated from whole colon tissue as described previously (Kim et al., 2018). Macrophages were further purified with MagniSort Mouse F4/80 Positive Selection Kit (Invitrogen, Carlsbad, CA, USA). Whole RNAs were extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA), followed by cDNA synthesis using a SuperScript II Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA). Gene expression was measured by StepOnePlus RT-PCR (Applied Biosystems, Waltham, MA, UNA). Primers used for PCR are listed in Supplementary Table 1. Each sample was run in triplicate and three independent experiments were performed for each sample.